Pro-drugs of NSAIAS with very high skin and membranes penetration rates and their new medicinal uses

ABSTRACT

The novel positively charged pro-drugs of NSAIAs in the general formulas (1, 2a, 2b, 2c, or 2d) “Structure 1, 2a, 2b, 2c, or 2d” were designed and synthesized. The compounds of the general formulas (1, 2a, 2b, 2c, or 2d) “Structure 1, 2a, 2b, 2c, or 2d” indicated above can be prepared from metal salts, organic base salts, or immobilized base salts of NSAIAs with suitable halide compounds. The positively charged amino groups in the pro-drugs in this invention largely increase the solubility of the drugs in water and will bond to the negative charge on the phosphate head group of membrane. Thus, the local concentration of the outside of the membrane or skin will be very high and will facilitate the passage of these pro-drugs from a region of high concentration to a region of low concentration. This bonding will disturb the membrane a little bit and may make some room for the lipophilic portion of the pro-drug. When the molecules of membrane move, the membrane may “crack” a little bit due to the bonding of the pro-drug. This will let the pro-drug insert into the membrane. At pH 7.4, only about 99% of the amino group is protonated. When the amino group is not protonated, the bonding between the amino group of the pro-drug and the phosphate head group of the membrane will disassociate, and the pro-drug will enter the membrane completely. When the amino group of the pro-drug flips to the other side of the membrane and thus becomes protonated, then the pro-drug is pulled into the cytosol, a semi-liquid concentrated aqueous solution or suspension. These pro-drugs can be used for treating and preventing diabetes (type I or/and type II), abnormal blood glucose and lipid levels, stroke, heart attack, and other heart and vascular diseases Alzheimer&#39;s diseases, Parkinson&#39;s diseases and other neurodegenerative diseases, psoriasis, discoid lupus erythematosus, systemic lupus erythematosus (SLE), autoimmune hepatitis, multiple sclerosis (MS), and other autoimmune diseases, amyotrophic lateral sclerosis (ALS), oculopharyngeal muscular dystrophy (OPMD), and other muscle disorders, inflamed hemorrhoids, cryptitis, other inflammatory conditions of the anorectum, and pruritus ani, prostatitis, prostatocystitis, varicose veins, autoimmune liver inflammation, autoimmune kidney inflammation, vein inflammation and other inflammations, skin cancers, breast cancer, colon-rectum cancer, oral cancer, and other cancers, scars, abnormal vascular skin lesions, birthmarks, moles (nevi), skin tags, aging spots (liver spots), and other skin disorders. These pro-drugs can be administered transdermally without the help of skin penetration enhancers.

TECHNICAL FIELD

The present invention relates to the design and preparation ofpositively charged and water-soluble pro-drugs of nonsteroidalanti-inflammatory agents (NSAIAs) with very high skin, scars,blood-milk, and brain-blood barriers penetration rates and their newmedicinal uses in treating and preventing diabetes (type I & II),abnormal blood glucose and lipid levels, stroke, heart attack, and otherheart and vascular diseases, Alzheimer's diseases, Parkinson's diseasesand other neurodegenerative diseases, psoriasis, discoid lupuserythematosus, systemic lupus erythematosus (SLE), autoimmune hepatitis,scleroderma, Sjogren's syndrome, rheumatoid arthritis, polymyositis,scleroderma, Hashimoto's thyroiditis, juvenile diabetes mellitus,Addison disease, vitiligo, pernicious anemia, glomerulonephritis, andpulmonary fibrosis, multiple sclerosis (MS), Crohn's disease, and otherautoimmune diseases, amyotrophic lateral sclerosis (ALS),oculopharyngeal muscular dystrophy (OPMD), myotonic dystrophy (MD),Duchenne muscular dystrophy (DMD), polymyositis (PM), dermatomyositis(DM), inclusion body myositis (IBM), and other muscle disorders,hemorrhoids, inflamed hemorrhoids, post irradiation (factitial)proctitis, chronic ulcerative colitis, cryptitis, other inflammatoryconditions of the anorectum, and pruritus ani, prostatitis,prostatocystitis, varicose veins, autoimmune liver inflammation,autoimmune kidney inflammation, colon-rectum inflammation, intestineinflammation, vein inflammation, vascular inflammation, and otherinflammations, skin cancers, breast cancer, colon-rectum cancer, oralcancer, lung and other respiratory system cancers, uterus cancer,genital cancer, urinary organs cancers, leukemia and other blood andlymph tissues cancers and other cancers, scars, abnormal vascular skinlesions, birthmarks, moles (nevi), skin tags, aging spots (liver spots),and other skin disorders. These pro-drugs can be administeredtransdermally without the help of skin penetration enhancers.

BACKGROUND ART

NSAIAs are used for the relief of signs and symptoms of rheumatoidarthritis, osteoarthritis and ankylosing spondylitis. NSAIAs are usedalone or as an adjunct in the treatment of biliary colic, fever, andepisiotomy pain. It is also used in treatment of gout, acute migraineheadaches, and renal colic and in the treatment of postoperativeinflammation in patients who have undergone cataract extraction. Aspirinis used for preventing heart and vascular diseases.

Unfortunately, a number of side effects are associated with the use ofNSAIAs, most notably GI disturbances such as dyspepsia, gastroduodenalbleeding, gastric ulcerations, and gastritis. Fishman (Fishman; Robert,U.S. Pat. No. 7,052,715) indicated that an additional problem associatedwith oral medications, is that the concentration levels which must beachieved in the bloodstream must be significant in order to effectivelytreat distal areas of pain or inflammation. These levels are often muchhigher than would be necessary if it were possible to accurately targetthe particular site of pain or injury. Fishman and many others (VanEngelen et al. U.S. Pat. No. 6,416,772; Macrides et al. U.S. Pat. No.6,346,278; Kirby et al. U.S. Pat. No. 6,444,234, Pearson et al. U.S.Pat. No. 6,528,040, and Botknecht et al. U.S. Pat. No. 5,885,597) haveattempted to develop a delivery system for transdermal application byformulation. Song, et al. developed a transdermal drug delivery systemfor anti-inflammatory analgesic agent comprising of diclofenacdiethylammonium salt (Song, et. al., U.S. Pat. No. 6,723,337). Donati,et al. developed a plaster for topical use containing heparin anddiclofenac. (Donati, et al., U.S. Pat. No. 6,592,891). Kawaji, et al.developed an oily patch for external use containing diclofenac sodium(Kawaji, et al. U.S. Pat. No. 6,262,121). Effing, et al. developed adevice for the transdermal delivery of diclofenac (Effing, et al. U.S.Pat. No. 6,193,996). It is very difficult, however, to delivertherapeutically effective plasma levels of NSAIAs into the host byformulation. Susan Milosovich, et. al. designed and preparedtestosteronyl-4-dimethylaminobutyrate.HCl (TSBH), which has a lipophilicportion and a tertiary amine groups that exists in the protonated format physiological pH. They found that the prodrug (TSBH) diffuses throughhuman skin ˜60 times faster than does the drug (TS) itself [SusanMilosovich, et al., J. Pharm. Sci., 82, 227 (1993).

DISCLOSURE OF INVENTION Technical Problem

Some NSAIAs have been used medicinally for more than 100 years. NSAIAsare indicated for the relief of the signs and symptoms of rheumatoidarthritis and osteoarthritis, the relief of mild to moderate pain, thereduction of fever, and the treatment of dysmenorrhea. They are the mostwidely used drugs in the world.

Unfortunately, a number of side effects are associated with the use ofNSAIAs, most notably GI disturbances such as dyspepsia, heartburn,vomiting, gastroduodenal bleeding, gastric ulcerations, and gastritis.Gastroduodenal bleeding induced by NSAIAs is generally painless but canlead to fecal blood loss and may cause a persistent iron deficiencyanemia.

Transdermal delivery systems help to avoid directly hurting thegastro-intestinal tract and inactivation of the drugs caused by the“first pass metabolism” in the gastro-intestinal tract and liver.Traditional transdermal drug delivery by using skin penetration enhancerhas limits. First, the penetration rates are very low (in μg/cm²/hscale). Second, large amounts of enhancer will enter the host body thatmay cause very serious side effects.

Technical Solution

Transdermal delivery systems help to avoid directly hurting thegastro-intestinal tract and inactivation of the drugs caused by the“first pass metabolism” in the gastro-intestinal tract and liver.Traditional transdermal drug delivery by using skin penetrationenhancers has limits. First, the penetration rates are very low (inμg/in²/h scale). Second, large amounts of enhancer will enter the hostbody and may cause extra side effects. Third, in traditional transdermaldrug delivery by using skin penetration enhancers, the high concentratedenhancers in the formulation may help the drug cross the skin, but whenthe enhancers and drug enter the skin, the concentration of enhancerswill be diluted greatly and they cannot provide any more help for drugmolecule to cross more biologic membranes, and then the drug moleculewill be accumulated in the fat layer under the skin and the accumulateddrugs may cause very serious and even fatal side effects.

Biological availability of a drug is the measurement of the relativeamount of administered drug that reaches the general circulation.However, the general circulation is not the “site of action” for most ofdrugs. Even if the drug molecules have reached the general circulation,they must cross more biologic membranes, which may be less permeablethan the gastrointestinal membranes, and interact with intercellular andintracellular fluids before reaching the elusive region called the “siteof action”; thus most drugs will be metabolized by intestinal mucosa,liver, blood, kidneys, and lungs before they reach the “site of action.”The situation not only produces very low pharmacological effect, butalso causes toxic burden on intestinal mucosa, blood, liver, kidneys,and lungs. If we can increase various membranes penetration rates ofdrugs, the pharmacological effect and the clinical response of drugswill be increased greatly, then a smaller drug dosage will be needed andfewer side effects will be caused. The transdermal pro-drug delivery forthe pro-drugs with very high skin and membranes penetration rates willbe very useful not only for local diseases, but also for systemicdiseases. Because these prodrugs have tens or hundreds of times morepotency than the parent drugs, only a few tenths or hundredths of thenormal drug dosage is needed and much less side effects will be caused.This will benefit not only transdermal drug delivery, but also to anyother drug delivery systems (such as oral, subcutaneous, intravenous,inhalation, and nasal).

We found that drugs that have a lipophilic portion and a primary,secondary, or tertiary amine group (preferably tertiary amine groups)that exists in the protonated form (hydrophilic portion) atphysiological pH can penetrate skin, scar, blood-brain, and blood-milkbarriers in very high rates (in mg/cm²/h scale). The principles fordesigning these prodrugs of NSAIAs are:

-   1. The prodrug must have a lipophilic portion and a primary,    secondary, or tertiary amine group (preferably tertiary amine    groups) that exists in the protonated form (hydrophilic portion) at    physiological pH.-   2. Every prodrug of NSAIAs should have only one or two (preferably    one) primary, secondary, or tertiary amine groups that exist in the    protonated form (hydrophilic portion) at physiological pH.-   3. The main role of the primary, secondary, or tertiary amine group    is to help the drug pass through the skin, scars, membrane,    blood-brain, blood-milk, and other barriers. The primary, secondary,    or tertiary amine group can be any structure that is non-toxic and    does not interfere the parent drugs' biologic activities.

We have disclosed some of the pro-drugs of all NSAIAs that have thegeneral formula (1) “Structure 1” in patents (international applicationNos: PCT/IB2006/052732, PCT/IB2006/052318, PCT/IB2006/052815,PCT/IB2006/052563, PCT/IB2006/052575, PCT/IB2006/053741,PCT/IB2006/053091, PCT/IB2006/053090, PCT/IB2006/052549).

In which, R represents a branched or straight chain —(CH₂)_(n)—, n=0, 1,2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, . . . , in —(CH₂)_(n)—, any CH₂ maybe replaced with O, S, NR₈, CH═CH, C≡C, CHR₈, CR₈R₉, aryl or heteroarylresidues, or any other moieties which are pharmaceutically acceptable;R₁ or R₂ represents H, one of any alkyl, alkyloxyl, alkenyl,perfluoroalkyl, alkyl halide or alkynyl residues having 1 to 12 carbonatoms, aryl or heteroaryl moieties, wherein, any CH₂ may be replacedwith O, S, CH═CH, C≡C, CHR₈, CR₈R₉, aryl or heteroaryl moieties, or anyother moieties which are pharmaceutically acceptable; X represents O,NH, NR₈, S, or none; R₈ represents H, OH, Cl, F, Br, I, one of anyalkyl, alkyloxyl, alkenyl, perfluoroalkyl, alkyl halide or alkynylresidues having 1 to 12 carbon atoms, aryl or heteroaryl moieties; R₉represents H, OH, Cl, F, Br, I, one of any alkyl, alkyloxyl, alkenyl,perfluoroalkyl, alkyl halide or alkynyl residues having 1 to 12 carbonatoms, aryl or heteroaryl moieties; HA represents none, HCl, HBr, HF,HI, HOAc, citric acid, or any acids which are pharmaceuticallyacceptable. All R, R₁, R₂, R₈, R₉ or —(CH₂)_(n)— groups are branched orstraight chains and may include C, H, O, Cl, Br, F, I, P, S, N or anyother atoms which are pharmaceutically acceptable and may have single,double, or/and triple bonds; all R, R₁, R₂, R₈, R₉ or —(CH₂)_(n)— groupsmay be achiral or chiral, if a group is chiral, it may have one or morechiral centers and may be a single (R) or (S) enantiomer or a mixture of(R) and (S) enantiomers; Ary- represents, but is not limited to:

wherein, R_(x) represents H, CH₃, CH₃O, OH, CH₃CH₂, CF₃, CHF₂, CH₂F, Cl,F, Br, F; R_(y) represents H, one of any alkyl, alkyloxyl, alkenyl,perfluoroalkyl, alkyl halide or alkynyl residues having 1 to 12 carbonatoms, aryl or heteroaryl moieties; X₁ or X₄ represents CH₂, S, O, NH,or CO; X₂ or X₅ represents CH, CR₈, or N; X₃ represents O, S, NH, orNR₈; Y, Y₁, Y₂, Y₃, Y₄, Y₅, or Y₆ represents independently H, HO,CH₃COO, R_(y)COO, HS, NO₂, CN, CH₃COS, NH₂, CH₃CONH, R_(y)CONH, CH₃,CH₃CH₂, C₃H₇, C₄H₉, CH₃O, CH₃CH₂O, C₃H₇O, Cl, F, Br, I, CH₃S, CHF₂O,CF₃O, CF₃CF₂O, C₃F₇O, CF₃, CF₃CF₂, C₃F₇, C₄F₉, CH₃SO₂, R_(y)SO₂, CH₃SO,R_(y)SO, CH3CO, CH₃CH₂CO; any Ary- may be achiral or chiral; If a Ary-is chiral, it may have one or more chiral centers and may be a single(R) or (S) enantiomer or a mixture of (R) and (S) enantiomers.

We found that the role of the primary, secondary, or tertiary aminegroup is only to help the drug pass through the skin, membrane,blood-brain, blood-milk, and other barriers, so the primary, secondary,or tertiary amine group can be any kind of structures which arenon-toxic and do not interfere the parent drugs' biologic activities. Sowe designed and prepared different kinds of amines groups for thisproperty. The new pro-drugs of NSAIAs have the general formulas (2a, 2b,2c, or 2d) “Structure 2a, 2b, 2c, or 2d”

wherein, R represents a branched or straight chain —(CH₂)_(n)—, whereinn=0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 . . . , in —(CH)₆—, any CH₂ may bereplaced with O, S, CH═CH, C≡C, CHR₆, CR₆R₇, aryl or heteroarylresidues, or other ring systems; R₁ represents a branched or straightchain, —(CH₂)_(a), wherein a=0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 . . . , in—(CH₂)_(a)—, any CH₂ may be replaced with O, S, CH═CH, C≡C, CHR₆, CR₆R₇,aryl or heteroaryl residues, or other ring systems; R₂ represents abranched or straight chain —(CH₂)_(b)—, wherein b=0, 1, 2, 3, 4, 5, 6,7, 8, 9, 10 . . . , in —(CH₂)_(b)—, any CH₂ may be replaced with O, S,CH═CH, C≡C, CHR₆, CR₆R₇, aryl or heteroaryl residues, or other ringsystems; R₃ represents a branched or straight chain, —(CH₂)_(c)—,wherein c=0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 . . . , in —(CH₂)_(c)—, anyCH₂ may be replaced with O, S, CH═CH, C≡C, CHR₆, CR₆R₇, aryl orheteroaryl residues, or other ring systems; R₄ represents H, one of anyalkyl, alkyloxy, alkenyl, perfluoroalkyl, alkyl halide, or alkynylresidues, having 1 to 12 carbon atoms, aryl or heteroaryl moieties,wherein, any CH₂ may be replaced with O, S, CH═CH, C≡C, CHR₆, CR₆R₇,aryl or heteroaryl moieties, or other ring moieties; R₅ represents H,one of any alkyl, alkyloxy, alkenyl, perfluoroalkyl, alkyl halide, oralkynyl residues, having 1 to 12 carbon atoms, aryl or heteroarylmoieties, wherein, any CH₂ may be replaced with O, S, CH═CH, C≡C, CHR₆,CR₇R₆, aryl or heteroaryl moieties, or other ring moieties; R₆represents H, one of any alkyl, alkyloxy, alkenyl, perfluoroalkyl, alkylhalide, or alkynyl residues having 1 to 12 carbon atoms, aryl orheteroaryl moieties, wherein, any CH may be replaced with O, S, CH═CH,C≡C, CHR₇, CR₇R₅, aryl or heteroaryl moieties, or other ring moieties,R₇ represents H, one of any alkyl, alkyloxy, alkenyl, perfluoroalkyl,alkyl halide, or alkynyl residues having 1 to 12 carbon atoms, aryl orheteroaryl moieties, wherein, any CH₂ may be replaced with O, S, CH═CH,C≡C, CHR₆, CR₆R₅, aryl or heteroaryl moieties, or other ring moieties; Xrepresents none, O, NH, NR₆, S; Ary- in the general formula (2a, 2b, 2c,or 2 d) “Structure 2a, 2b, 2c, or 2d” are defined as same Ary- as in thegeneral formula (1) “Structure 1”; HA represents none, HCl, HBr, HF, HI,HOAc, citric acid, or any acids which are pharmaceutically acceptable.All R, R₁, R₂, R₃, R₄, R₅, R₆, R₇, or —(CH₂)_(n)— groups are branched orstraight chains and may include C, H, O, Cl, Br, F, I, P, S, N or anyother atoms which are pharmaceutically acceptable and may have single,double, or/and triple bonds; all R, R₁, R₂, R₃, R₄, R₅, R₆, R₇, or—(CH₂)_(n)— groups may be achiral or chiral, if a group is chiral, itmay have one or more chiral centers and may be a single (R) or (S)enantiomer or a mixture of (R) and (S) enantiomers.

Drug absorption, whether from the gastrointestinal tract or other sites,requires the passage of the drug in a molecular form across the barriermembrane. The drug must first dissolve, and if the drug possesses thedesirable biopharmaceutical properties, it will pass from a region ofhigh concentration to a region of low concentration across the membraneinto the blood or general circulation. All biological membranes containlipids as major constituents. The molecules that play the dominant rolesin membrane formation all have phosphate-containing highly polar headgroups, and, in most cases, two highly hydrophobic hydrocarbon tails.Membranes are bilayers, with the hydrophilic head groups facing outwardinto the aqueous regions on either side. Very hydrophilic drugs cannotpass the hydrophobic layer of membrane and very hydrophobic drugs willstay in the hydrophobic layer as part of the membrane due to theirsimilarities and cannot enter the cytosol on the inside efficiently.

One goal of this invention is to avoid the side effects of NSAIAs byincreasing the solubility of NSAIAs in gastric juice which will make itadministrable orally and the penetration rate of NSAIAs through themembranes and skin barrier which will make it administrabletransdermally (topical application). The most important goal of thisinvention is to design pro-drugs of NSAIAs that can penetrate the skin,cell membrane, especially the brain cell and nerve cell membranes, veryeffectively and stay the general circulation much shorter, thus theywill have tens or hundreds of times more potency than the parent drugs,only a few tenths or hundredths of the normal drug dosage is needed andmuch less side effects will be caused. This will benefit not onlytransdermal drug delivery, but also any other drug delivery systems(such as oral, subcutaneous, intravenous, inhalation, and nasal) and cantreat many conditions that cannot be treated by their parent drugs.These novel pro-drugs of NSAIAs have two structural features in common:they have a lipophilic portion and a primary, secondary, or tertiaryamine group that exists in the protonated form (hydrophilic part) atphysiological pH. Such a hydrophilic-lipophilic balance is required forefficient passage through the membrane barrier [Susan Milosovich, etal., J. Pharm. Sci., 82, 227 (1993)]. The positively charged aminogroups largely increase the solubility of the drugs in water. Thepositive charge on the amino groups of these pro-drugs will bond to thenegative charge on the phosphate head group of membrane. Thus, the localconcentration of the outside of the membrane or skin will be very highand will facilitate the passage of these pro-drugs from a region of highconcentration to a region of low concentration. This bonding willdisturb the membrane a little bit and may make some room for thelipophilic portion of the pro-drug. When the molecules of the membranemove, the membrane may “crack” a little bit due to the bonding of thepro-drug. This will let the pro-drug insert into the membrane. At pH7.4, only about 99% of the amino group is protonated. When the aminogroup is not protonated, the bonding between the amino group of theprodrug and the phosphate head group of the membrane will disassociate,and the pro-drug will enter the membrane completely. When the aminogroup of the pro-drug flips to the other side of the membrane and thusbecomes protonated, then the pro-drug is pulled into the cytosol, asemi-liquid concentrated aqueous solution or suspension. Due to theshort stay in GI tract, the pro-drugs will not cause gastric mucosalcell damage. The penetration rates of the novel pro-drugs through humanskin were measured in vitro by using modified Franz cells, which wereisolated from human skin tissue (360-400 μm thick) of the anterior andposterior thigh areas. The receiving fluid consisted of 10 ml of 2%bovine serum albumin in normal saline and was stirred at 600 rpm. Thecumulative amounts of pro-drugs and drugs entering the skin versus timewere determined by a specific high-performance liquid chromatographymethod. Apparent flux values of the pro-drugs of NSAIAs are 0.1-50mg/cm²/h. The results suggest that the pro-drug diffuses through humanskin at least a hundred times faster than do their parent drugs. Theresults suggest that the positive charge on the dialkyaminoethyl grouphas a very important role in the passage of the drug across the membraneand skin barrier.

The novel prodrugs of NSAIAs can penetrate the skin barrier, blood-brainbarrier, and blood-milk barrier. The in vivo rates of penetration ofpro-drugs and their parent drugs through the skin of intact rats werecompared. The donor consisted of 20% pro-drugs or their parent drugs in1 ml of ethanol applied to about 5 cm² area on the backs of rats (˜200g). After 4 hours, the rats were killed, and 5 ml of methanol was addedto 1 ml of blood, 1 g of liver, 1 g of kidney, or 1 g of brain (liver,kidney or brain was washed with pH 7.2 buffer for three times) and themixtures were homogenized. The samples were then centrifuged for 5 minand analyzed using HPLC. The results are showed in table 1-5.

TABLE 1 The distribution of diethylaminoethylacetylsalicylate•acetylsalicylic acid salt (P-1) and its metabolites inrats' body tissues and plasma. Prodrug or metabolites Plasma LiverKidney Brain P-1 30 ± 10 μg/ml 15 ± 8 μg/g 25 ± 6 μg/g 15 ± 6 μg/gAspirin 25 ± 8 μg/ml  13 ± 8 μg/g 20 ± 6 μg/g 15 ± 5 μg/g Salicylic 80 ±10 μg/ml 30 ± 8 μg/g 45 ± 6 μg/g 3 0 ± 6 μg/g  acid

TABLE 2 The distribution of 1-piperidinepropyl2[(2,6-dichlorophenyl)amino]benzene acetate•AcOH (P-2) and itsmetabolite in rats' body tissues and plasma. Prodrug or metabolitePlasma Liver Kidney Brain P-2 40 ± 10 μg/ml 22 ± 8 μg/g 20 ± 6 μg/g 25 ±6 μg/g diclofenac 75 ± 8 μg/ml  25 ± 8 μg/g 4 8 ± 6 μg/g  40 ± 5 μg/g

TABLE 3 The distribution of 1-pyrrolidinepropyl 2-(3-benzoylphenyl)propionate•AcOH (P-3) and its metabolite in rats' body tissues andplasma. Prodrug or metabolite Plasma Liver Kidney Brain P-3 35 ± 8 μg/ml22 ± 8 μg/g 25 ± 6 μg/g 20 ± 6 μg/g ketoprofen 70 ± 8 μg/ml 32 ± 8 μg/g45 ± 6 μg/g 35 ± 5 μg/g

TABLE 4 The distribution of 4-piperidinemethyl2-(3-phenoxyphenyl)propionate•AcOH (P-4) and its metabolite in rats'body tissues and plasma. Prodrug or metabolite Plasma Liver Kidney BrainP-4 32 ± 8 μg/ml 20 ± 8 μg/g 20 ± 6 μg/g 20 ± 6 μg/g fenoprofen 80 ± 8μg/ml 38 ± 8 μg/g 48 ± 6 μg/g 45 ± 5 μg/g

TABLE 5 The distribution of 3-piperidinemethyl 2-(ρ-isobutylphenyl)propionate•AcOH (P-5) and its metabolite in rats body' tissues andplasma. Prodrug or metabolite Plasma Liver Kidney Brain P-5 40 ± 8 μg/ml25 ± 8 μg/g 30 ± 6 μg/g 25 ± 6 μg/g ibuprofen 70 ± 8 μg/ml 35 ± 8 μg/g45 ± 6 μg/g 35 ± 5 μg/g

Then 20% of aspirin, diclofenac, ketoprofen, fenoprofen, or ibuprofen in1 ml of ethanol applied to about 5 cm² area on the backs of rats. After4 hours, the rats were killed, and 5 ml of methanol was added to 1 ml ofblood, 1 g of liver, 1 g of kidney, or 1 g of brain (liver, kidney orbrain was washed with pH 7.2 buffer for three times) and the mixtureswere homogenized. None of these drugs was found in any rat's tissues orplasma. The results show that the pro-drugs of NSAIAs can penetrate theskin barrier, blood-brain barrier, and other membrane barriers at a veryhigh rate, but the parent NSAIAs cannot penetrate the skin barrier in adetectable amount.

The pro-drugs of the general formula (1) “Structure 1” have demonstratedanti-inflammatory, analgesic, antipyretic, and antirheumatic activity inour patents (international application Nos: PCT/IB2006/052732,PCT/IB2006/052318, PCT/IB2006/052815, PCT/IB2006/052563,PCT/IB2006/052575, PCT/IB2006/053741, PCT/IB2006/053091,PCT/IB2006/053090, PCT/IB2006/052549). We found that the pro-drugs ofthe general formula (2a, 2b, 2c, or 2d) “Structure 2a, 2b, 2c, or 2d”,have demonstrated anti-inflammatory, analgesic, antipyretic, andantirheumatic activity. The main focuses of this invention are the newmedicinal uses of the pro-drugs of NSAIAs.

The relationship between inflammation and cancer is well known. Dr. TheaD. Tlsty described in his speech (Keystone Symposia: Inflammation andCancer, Breckenridge, Colo., USA, Feb. 27-Mar. 3, 2005) thatcyclooxygenase-2 (COX-2) stimulates aromatase activity, angiogenesis,proliferation, invasion, and prostaglandin synthesis. The increase inprostaglandins leads to an inhibition of apoptosis. Aspirin and otherNSAIAs inhibit COX-1 and COX-2. The overall relative risk of colorectalcancer, oesophageal cancer, ovarian cancer or other cancers is reducedin people taking long term aspirin. However, cancer cells may changetheir membrane structure to keep the NSAIAs from entering the cancercells. The novel pro-drugs in this invention can penetrate any membranebarriers and can be applied topically to the outside skin area of thelocation of the cancer and large amounts of the pro-drugs will enter thecancer cells with very little systemic exposure.

For evaluation of anti-tumor activity of the pro-drugs of NSAIAs, humanbreast cancer cells (BCAP-37, 2-3 mm³ of tumor tissue was used in eachmouse) were subcutaneously xenografted into nude mice (BALB, 12 groups,7 mice each group). After 14 days, the tumors were growing to the sizeof 50±10 mm³ (0.05 ml). Then 30 μl of 5% (equal to 1.5 mg of thepro-drugs) diethylaminoethyl acetylsalicylate.acetylsalicylic acid salt(P-1, in acetone); 1-piperidinepropyl2[(2,6-dichlorophenyl)amino]benzene acetate.AcOH (P-2, in water),1-pyrrolidinepropyl 2-(3-benzoylphenyl)propionate.AcOH (P-3, in water),4-piperidinemethyl 2-(3-phenoxyphenyl)propionate.AcOH (P-4, in water),3-piperidinemethyl 2-(ρ-isobutylphenyl)propionate.AcOH (P-5, in water),diethylaminoethyl 1-(ρ-chlorobenzoyl)-5-methoxy-2-methylindole3-acetate.AcOH (P-11, in water), 2-(4-morpholinyl)ethyl(Z)-5-fluoro-2-methyl-1-[(4-methylsulfinyl)phenylmethylene]-1H-indene-3-acetate.AcOH(P-12, in water), diethylaminoethyl2-(2,4-dichlorophenoxy)benzeneacetate.AcOH (P-19, in water),diethylaminoethyl2-(8-methyl-10,11-dihydro-11-oxodibenz(b,f)oxepin-2-yl)propionate.AcOH(P-37, in water), 1-pyrrolidinepropyl2-[[(3-(trifluoromethyl)phenyl)amino]benzoate.AcOH (P-48, in water),4-N,N-dimethylaminobutyryloxy-2-methyl-N-2-pyridinyl-2H,1,2-benzothiazine-3-carboxamide 1,1-dioxide.HCl (P-51, in acetone) was topically applied to thehuman breast cancer cells-implanted area (near the front leg) every 8hours. At day 42, the tumors sizes are shown in table 6 and table 7.

Table 6, the tumors sizes and the weights of the control group and thedrug-treated groups of nude mice at day 42.

TABLE 6 Pro-drug Control P-1 P-2 P-3 P-4 P-5 Size (mm³) 800 ± 100 150 ±50 180 ± 50 200 ± 50 180 ± 50 190 ± 50 Weight 22 ± 2  22 ± 3 22 ± 2 21 ±3 22 ± 3 23 ± 2

Table 7, the tumors sizes and the weights of the drug-treated groups ofnude mice at day 42.

TABLE 7 Pro-drug P-11 P-12 P-19 P-37 P-48 P-51 Size (mm³) 210 ± 100 250± 50 280 ± 50 250 ± 50 290 ± 50 390 ± 50 Weight 21 ± 2  23 ± 3 21 ± 2 23± 3 22 ± 3 23 ± 2

In the second anti-tumor experiment, human colon cancer cells (LS174J,2-3 mm³ of tumor tissue was used in each mouse) were subcutaneouslyxenografted into nude mice (BALB). After 7 days, the tumors were growingto the size of 55±10 mm³ (0.055 ml). Then about 30 μl of 5% (equal to1.5 mg of the pro-drugs) diethylaminoethylacetylsalicylate.acetylsalicylic acid salt (P-1, in acetone);1-piperidinepropyl 2[(2,6-dichlorophenyl)amino]benzene acetate.AcOH(P-2, in water), 1-pyffolidinepropyl 2-(3-benzoylphenyl)propionate.AcOH(P-3, in water), 4-piperidinemethyl 2-(3-phenoxyphenyl)propionate.AcOH(P-4, in water), 3-piperidinemethyl 2-(ρ-isobutylphenyl)propionate.AcOH(P-5, in water), diethylaminoethyl1-methyl-5-(4-methylbenzoyl)-1H-pyrrole-2-acetate.AcOH (P-13, in water),2-(4-morpholinyl)ethyl 2-amino-3-benzoylbenzeneacetate.AcOH (P-16, inwater), diethylaminoethyl2-(10,11-dihydro-10-oxodibenzo(b,f)thiepin-2-yl)propionate.AcOH (P-36),diethylaminoethyl 2-[(2,3-dimethylphenyl)amino]benzoate.AcOH (P-46, inwater), diethylaminoethyl2-[(2,6-dichloro-3-methylphenyl)amino]benzoate.AcOH (P-47, in water),N-(2-thiazoyl)-4-N,N-dimethylaminobutyryloxy-2-methyl-2H,1,2-benzothiazine-3-carboxamide1,1-dioxide.HCl (P-52, in acetone) was topically applied to the humancolon cancer cells-implanted area (near the front leg) every 8 hours. Atday 30, the tumors sizes are shown in table 8 and table 9.

Table 8, the tumors sizes and the weights of the control group and thedrug-treated groups of nude mice at day 30.

TABLE 8 Pro-drug Control P-1 P-2 P-3 P-4 P-5 Size (mm³) 1300 ± 300 420 ±100 480 ± 180 500 ± 150 480 ± 120 390 ± 110 Weight 21 ± 2 22 ± 3  22 ±2  21 ± 3  22 ± 3  23 ± 2 

Table 9, the tumors sizes and the weights of the drug-treated groups ofnude mice at day 30.

TABLE 9 Pro-drug P-13 P-16 P-36 P-46 P-47 P-52 Size (mm³) 610 ± 200 550± 150 480 ± 180 650 ± 250 490 ± 150 690 ± 250 Weight 21 ± 2  23 ± 3  21± 2  23 ± 3  22 ± 3  23 ± 2 

The results show that the pro-drugs of NSAIAs have very stronganti-tumor activity and have none or very little side effects.

The hypoglycemic effect of salicylates was first observed over 100 yearsago by German physicians (Edmund J. Hengesh, Principles of medicinalchemistry, 4th ed., pg 591, Williams & Wilkins, 1995). Salicylatesenhance glucose-stimulated insulin secretion and inhibits glucogenesisfrom lactate and alanine (H. F. Woods, et al., Clin. Exp. PharmacolPhysiol., 1, 534 (1974). Certain salicylates decrease plasma levels offree fatty acids, triglycerides, and cholesterol. Because elevatedlevels of plasma-free fatty acids inhibit glucose utilization, adecrease in their concentration could contribute to the hypoglycemicaction. Unfortunately, the large doses (5 g daily) of salicylates arenecessary to maintain adequate control of blood sugar levels and bloodlipid levels. At these dosage levels, numerous side effects, such asgastric irritation, nausea, vomiting, and tinnitus, occur frequently.The novel pro-drugs in this invention have very high skin and membranepenetration rates. They can reach the “site of action” very fast and thepharmacologic effect and the clinical response of these pro-drugs areincreased greatly, then much smaller (only hundredths to tenths of theparent drug dosage needed) drug dosage will be needed and much less sideeffects will be caused.

The pro-drugs in this invention lower blood glucose levels in rat models(SLAC/GK, type 2 diabetes, n=7). 50% acetone solution ofdiethylaminoethyl acetylsalicylate.acetylsalicylic acid salt (P-1, inacetone); 4-acetamidophenyl salicylyldimethylaminobutyrate.HCl (P-6),diethylaminoethyl 5-(2,4-difluorophenyl)acetylsalicylate.5-(2,4-difluorophenyl)acetylsalicylic acid salt (P-8), diethylaminoethylsalicylsalicylate.AcOH (P-9), diethylaminoethyl salicylate.AcOH (P-10),diethyllaminoethyl 5-acetamido-acetylsalicylate (P-58),diethylaminoethyl acetylsalicylsalicylate.acetylsalicylsalicylic acidsalt (P-59), diethylaminoethylacetylsalicylsalicylate.acetylsalicylsalicylsalicylic acid salt (P-60)(equal to of 20 mg/kg of NSAIAs) were administered transdermally to thebacks (about 1.5 cm²) of rats (fur was shaved) once per day (at 8 am)for 5 weeks. The blood glucose levels were measured once every 3 days at4 pm (no fasting) from the second week to the fifth week. The resultsare shown in table 10. The blood lipid levels were measured at the endof the fifth week. The results are shown in table 11.

TABLE 10 Anti-diabetes activity of the pro-drugs of NSAIAs ProdrugControl P-1 P-6 P-8 P-9 P-10 P-58 P-59 P-60 mmol/L mmol/L mmol/L mmol/Lmmol/L mmol/L mmol/L mmol/L mmol/L Diabetic Baseline 15.6 ± 3  16.1 ± 3 16.7 ± 4  17.1 ± 3  16.5 ± 4  15.8 ± 3  17.1 ± 3  16.3 ± 3  15.5 ± 3 rats Average 15.9 ± 3  6.5 ± 1 8.5 ± 2 8.1 ± 1 8.4 ± 1 8.2 ± 1 8.4 ± 18.7 ± 1 8.6 ± 1 Normal Baseline 6.5 ± 1 6.4 ± 1 6.8 ± 1 7.1 ± 1 6.5 ± 16.8 ± 1 6.9 ± 1 7.2 ± 1 6.6 ± 1 rats Average 6.6 ± 1 6.3 ± 1 6.5 ± 1 6.8± 1 6.7 ± 1 6.9 ± 1 7.1 ± 1 7.3 ± 1 7.5 ± 1

The results showed that the pro-drug s of NSAIAs lowered blood glucoselevels in diabetes rat models very effectively and did not affect theblood glucose levels of normal rats. The most interesting thing is thatthe blood glucose levels of the rats still stayed at normal levels (7-8mmol/L, no fasting) after the treatment was stopped for 30 days. Thismeans that the pro-drugs not only control blood glucose levels, but alsomay cure diabetes.

TABLE 11 Blood lipid-lowering activity of the pro-drugs of NSAIAsProdrug Control P-1 P-6 P-8 P-9 P-10 P-58 P-59 P-60 mmol/L mmol/L mmol/Lmmol/L mmol/L mmol/L mmol/L mmol/L mmol/L cholesterol Baseline 7.6 ± 0.57.7 ± 0.4 7.3 ± 0.5 7.6 ± 0.6 7.7 ± 0.5 7.1 ± 0.5 7.8 ± 0.5 7.6 ± 0.67.3 ± 0.6 (total) Average 7.9 ± 0.5 4.0 ± 0.3 4.7 ± 0.4 5.3 ± 0.3 4.8 ±0.4 4.9 ± 0.4 5.4 ± 0.4 5.1 ± 0.3 5.2 ± 0.3 Cholesterol Baseline 1.4 ±0.1 1.4 ± 0.2 1.3 ± 0.1 1.6 ± 0.2 1.3 ± 0.1 1.6 ± 0.2 1.3 ± 0.1 1.6 ±0.2 1.3 ± 0.2 (HDL) Average 1.3 ± 0.1 1.5 ± 0.2 1.3 ± 0.2 1.5 ± 0.2 1.4± 0.2 1.5 ± 0.1 1.4 ± 0.2 1.6 ± 0.2 1.5 ± 0.2 Triglycerides Baseline 5.2± 0.7 5.9 ± 0.5 6.3 ± 0.5 5.6 ± 0.6 5.7 ± 0.5 6.6 ± 0.6 5.3 ± 0.5 5.6 ±0.5 5.7 ± 0.6 Average 5.5 ± 0.6 1.5 ± 0.2 2.3 ± 0.2 2.1 ± 0.2 2.6 ± 0.22.9 ± 0.2 2.4 ± 0.2 2.9 ± 0.2 2.5 ± 0.2

The results showed that the pro-drug s of NSAIAs lowered blood lipidlevels (total cholesterol and triglycerides) in diabetes rat models veryeffectively and did not affect HDL levels.

The pH of stomach juice is 1-3. The negative charge on the phosphatehead group of membrane is neutralized by the proton and the positivecharge on the amino groups of these pro-drugs cannot bond to thephosphate head group of membranes, then the pro-drugs cannot passthrough the wall of the stomach and will not hurt or upset the stomach.The pH of the duodenum is about 5-7 and the pro-drugs can pass throughthe mucosa of duodenum. The pancreas is nearby and large amount of thepro-drugs will enter there before going to the liver, kidneys, and thegeneral circulation where the drug will be metabolized, so only very lowdose of these pro-drugs are needed and very few and low side effectswill be caused. The 20% acetone solution of diethyllaminoethylacetylsalicylate.acetylsalicylic acid salt (P-1, in acetone);4-acetamidophenyl salicylyldimethylaminobutyrate.HCl (P-6),diethylaminoethyl 5-(2,4-difluorophenyl)acetylsalicylate.5-(2,4-difluorophenyl)acetylsalicylic acid salt (P-8), diethylaminoethylsalicylsalicylate.AcOH (P-9), diethylaminoethyl salicylate.AcOH (P-10),diethylaminoethyl 5-acetamido-acetylsalicylate (P-58), diethylaminoethylacetylsalicylsalicylate.acetylsalicylsalicylic acid salt (P-59),diethylaminoethyl acetylsalicylsalicylate.acetylsalicylsalicylsalicylicacid salt (P-60) (equal to of 15 mg/kg of NSAIAs) were mixed with foodand were orally administered to the rats (SLAC/GK, type 2 diabetes, n=7)with food every day for 5 weeks. The blood glucose levels were measuredonce every 3 days at 3 pm (no fasting) from the second week to the fifthweek. The results are shown in table 12. The blood lipid levels weremeasured at the end of the fifth week. The results are shown in table13.

TABLE 12 Anti-diabetes activity of the pro-drugs of NSAIAs ProdrugControl Control P-1 P-6 P-8 P-9 P-10 P-58 P-59 P-60 mmol/L mmol/L mmol/Lmmol/L mmol/L mmol/L mmol/L mmol/L mmol/L mmol/L Diabetic Baseline 15.3± 3  16.5 ± 3  16.1 ± 4  16.1 ± 3  16.5 ± 4  15.6 ± 3  17.0 ± 3  15.3 ±3  16.5 ± 3  rats Average 15.6 ± 3  6.5 ± 1 7.5 ± 2 7.3 ± 1 7.6 ± 1 7.8± 1 8.4 ± 1 8.6 ± 1 7.9 ± 1 Normal Baseline 6.6 ± 1 6.3 ± 1 6.5 ± 1 7.0± 1 6.3 ± 1 6.7 ± 1 6.9 ± 1 7.5 ± 1 6.8 ± 1 rats Average 6.5 ± 1 6.5 ± 16.4 ± 1 6.8 ± 1 6.6 ± 1 6.9 ± 1 7.2 ± 1 7.3 ± 1 7.3 ± 1

The results showed that the pro-drug s of NSAIAs lowered blood glucoselevels in diabetic rat models very effectively and did not affect theblood glucose levels of normal rats when the pro-drugs were taken orallyand the dosages are much smaller than that of the parent drugs.

TABLE 13 Blood lipid-lowering activity of the pro-drugs of NSAIAsProdrugs Control P-1 P-6 P-8 P-9 P-10 P-58 P-59 P-60 mmol/L mmol/Lmmol/L mmol/L mmol/L mmol/L mmol/L mmol/L mmol/L Cholesterol Baseline7.8 ± 0.6 7.7 ± 0.4 7.5 ± 0.4 7.4 ± 0.6 7.9 ± 0.5 7.6 ± 0.5 7.9 ± 0.57.7 ± 0.6 7.5 ± 0.5 (total) Average 8.1 ± 0.5 4.1 ± 0.3 4.7 ± 0.4 5.1 ±0.3 5.3 ± 0.4 5.2 ± 0.5 4.9 ± 0.4 5.1 ± 0.3 5.0 ± 0.3 CholesterolBaseline 1.7 ± 0.1 1.5 ± 0.1 1.8 ± 0.1 1.4 ± 0.2 1.5 ± 0.1 1.6 ± 0.2 1.8± 0.1 1.9 ± 0.2 1.5 ± 0.2 (HDL) Average 1.5 ± 0.1 1.6 ± 0.2 1.4 ± 0.21.3 ± 0.2 1.4 ± 0.2 1.7 ± 0.1 1.6 ± 0.2 1.8 ± 0.2 1.6 ± 0.2Triglycerides Baseline 5.5 ± 0.6 5.7 ± 0.5 5.9 ± 0.5 6.2 ± 0.7 5.8 ± 0.65.7 ± 0.6 5.9 ± 0.5 5.4 ± 0.6 6.2 ± 0.5 Average 5.8 ± 0.6 1.4 ± 0.2 1.8± 0.2 1.8 ± 0.2 2.7 ± 0.2 2.6 ± 0.2 2.8 ± 0.2 2.7 ± 0.2 2.5 ± 0.2The results showed that the pro-drug s of NSAIAs lowered blood lipidlevels (total cholesterol and triglycerides) in diabetic rat models veryeffectively when the pro-drugs were taken orally and the dosages aremuch smaller than that of the parent drugs.

The pro-drugs in this invention lower blood glucose levels in mousemodels (SLAC:NOD-IDDM, type 1 diabetes, n=7). 20% acetone solution ofdiethylaminoethyl acetylsalicylate.acetylsalicylic acid salt (P-1, inacetone); 4-acetamidophenyl salicylyldimethylaminobutyrate.HCl (P-6),diethylaminoethyl5-(2,4-difluorophenyl)acetylsalicylate.5-(2,4-difluorophenyl)acetylsalicylicacid salt (P-8), diethylaminoethyl salicylsalicylate.AcOH (P-9),diethylaminoethyl salicylate.AcOH (P-10), diethyllaminoethyl5-acetamido-acetylsalicylate (P-58), diethylaminoethylacetylsalicylsalicylate.acetylsalicylsalicylic acid salt (P-59),diethylaminoethyl acetylsalicylsalicylate.acetylsalicylsalicylsalicylicacid salt (P-60) (equal to of 20 mg/kg of NSAIAs) were administeredtransdermally to the backs (about 1 cm²) of mice (fur was shaved) onceper day (at 8 am) for 7 weeks. The blood glucose levels were measuredonce every 3 days at 3 pm (no fasting) from the fourth week to theseventh week. The results are shown in table 14.

TABLE 14 Anti-diabetes (type I) activity of the pro-drugs of NSAIAsProdrug Control P-1 P-6 P-8 P-9 P-10 P-58 P-59 P-60 mmol/L mmol/L mmol/Lmmol/L mmol/L mmol/L mmol/L mmol/L mmol/L Baseline 28.6 ± 5 26.1 ± 527.7 ± 4 29.1 ± 5 26.5 ± 4 25.8 ± 3 27.1 ± 3 24.3 ± 3 25.5 ± 3 Average32.9 ± 5  6.5 ± 1  9.5 ± 2  9.1 ± 1  9.4 ± 1  8.2 ± 1  7.9 ± 1  8.7 ± 1 8.6 ± 1The results showed that the pro-drugs of NSAIAs lowered blood glucoselevels in diabetic (type I) mouse models very effectively.

Eighteen Chinese White rabbits weighing between 3.0 and 3.5 kg (aged 6-7months) were selected and divided into three groups (control, P-1 andP-10 groups, n=6). One hour before the experiment, thrombi were made byaspirating venous blood (1 ml) into a sterilized bottle to clot. Toavoid fragmentation and slow lysis, the autologous blood clots werestabilized in temperature-controlled (70° C.) distilled water for 10min. After anesthesia, the femoral veins were exposed and distallyisolated, and autologous blood clots (0.05 g/kg) were injected throughan indwelling catheter (20GA), which had been placed in the femoral veinisolated earlier. 50% acetone solution of diethylaminoethylacetylsalicylate.acetylsalicylic acid salt (P-1, in acetone, 15 mg/kg)and diethylaminoethyl acetylsalicylsalicylate.acetylsalicylsalicylicacid salt (P-59, 15 mg/kg) were topically applied to the back of therabbits. After 2 days, rabbits were euthanized with an excessiveintravenous injection of sodium amobarbital (60 mg/kg). The lungs andhearts were isolated to observe whether thrombi were present in thepulmonary arteries. The lungs were immersed in 10% formalin for 24 h.Consecutive transverse sections along the obstructed pulmonary arterieswere paraffin-embedded and stained with hematoxylin-eosine. In thecontrol group, platelet thrombus and mixed thrombus surrounded theinfused clots, which were present in large-sized vessels as well andstretched the vessel walls in both proximal and distal directions. Therewas excessive proliferation of endothelial cells and fibrocytes in thesevessels. Additionally, there was acute pulmonary congestion. In the P-1and P-59 groups, both lung tissue and vascular walls were normal. Theresults showed that thrombotic activity and that embolization-associatedthrombus propagation can be prevented by these pro-drugs of NSAIAs.These pro-drugs can be very useful for preventing and treating bloodclots—a major cause of strokes, heart attacks and organ transplantrejection.

The pro-drugs in this invention can help heal wounds and soften andshrink scars from cuts and burns in rabbit models. The average scar areaof the pro-drugs treated rabbits is only a third of that of the controlrabbits from same size cut wounds in the Chinese white rabbit model andthe scars are as soft as normal unscarred tissues.

COX-1 and COX-2 play a very important role in animal immune-responses.NSAIAs inhibite COX-1 and COX-2. The pro-drugs of NSAIAs in thisinvention may be very useful for treating psoriasis, discoid lupuserythematosus, systemic lupus erythematosus (SLE), and other autoimmunediseases. Heavy suspensions of Malassezia [Rosenberg, E. W., et al.,Mycopathologia, 72, 147-154 (1980)] were applied to the shaved skin onthe backs of the Chinese white rabbits (n=4×6) twice (at 7 am and 7 pm)per day for 2 weeks, lesions similar to psoriasis resulted. Then a 5%aqueous solution of 3-piperidinemethyl2-(ρ-isobutylphenyl)propionate.AcOH (P-5), diethyllaminoethyl1-methyl-5-(4-methylbenzoyl)-1H-pyrrole-2-acetate.AcOH (P-13),diethyllaminoethyl5-(4-Chlorobenzoyl)-1,4-dimethyl-1H-pyrrole-2-acetate.AcOH (P-14),diethylaminoethyl1,8-diethyl-1,3,4,9-tetrahydropyrano-[3,4-b]indole-1-acetate.AcOH(P-15), diethylaminoethyl 2-amino-3-(4-bromo-benzoyl)benzeneacetate.AcOH(P-17) diethylaminoethyl 3-chloro-4-(2-propenyloxy)benzeneacetate.AcOH(P-18), diethyllaminoethyl1-(4-chlorobenzoyl-5-methoxy-2-methyl-1H-indole-3-acetoxyacetate.AcOH(P-20), diethylaminoethyl4-(4-chlorophenyl)-2-phenyl-5-thiazoleacetate.AcOH (P-21),diethylaminoethyl 3-(4-chlorophenyl)-1-phenyl-1H-pyrazole-4-acetate.AcOH(P-22) were applied to the same areas 3 hours (10 am and 10 pm) afterthe application of heavy suspensions of Malassezia (7 am and 7 pm). 10days after the application of these pro-drugs, the lesions wereresolved. but the condition of the control mice were getting worse.

For evaluation of anti-lupus erythematosus activity, 5%diethylaminoethyl acetylsalicylate.acetylsalicylic acid salt (P-1, inacetone, 30 mg/kg) or 3-piperidinemethyl2-(ρ-isobutylphenyl)propionate.AcOH (P-5, in water, 30 mg/kg) weretopically applied to the skin on the backs of mice (MRL/LPR, n=5×3) withdiscoid lupus erythematosus and systemic lupus erythematosus twice perday. After 6 weeks, all skin lesions and lupus nephritis were resolvedin the pro-drug treated mice, but the condition of the control mice weregetting worse.

These results suggest that these pro-drugs of NSAIAs are very promisingagents for the treatment of psoriasis, discoid lupus erythematosus,systemic lupus erythematosus (SLE), multiple sclerosis (MS), and otherautoimmune diseases in human.

The pathogenesis of cell death in amyotrophic lateral sclerosis (ALS)may involve glutamate-mediated excitotoxicity, oxidative damage, andapoptosis. Cyclooxygenase-2, present in spinal neurons and astrocytes,catalyzes the synthesis of prostaglandin E2. Prostaglandin E2 stimulatesglutamate release from astrocytes, whereas cyclooxygenase-2 also plays akey role in the production of pro-inflammatory cytokines, reactiveoxygen species, and free radicals. Treatment with a selectivecyclooxygenase-2 inhibitor, celecoxib, markedly inhibited production ofprostaglandin E2 in the spinal cords of ALS mice. Celecoxib treatmentsignificantly delayed the onset of weakness and weight loss andprolonged survival by 25%. Spinal cords of treated ALS mice showedsignificant preservation of spinal neurons and diminished astrogliosisand microglial activation (Merit. E. Cudkowicz, et al., Annals ofneurology, 52, 771-778, 2002). These results suggest thatcyclooxygenase-2 inhibition may benefit ALS patients. The pro-drugs ofNSAIAs in this invention can penetrate skin and nerve cell membranebarriers in very high rates (most NSAIAs cannot penetrate nerve cellseffectively) and can be administered transdermally without hurting theGI tract, so these pro-drug are very promising agents for the treatmentof multiple sclerosis (MS), Crohn's disease, and other autoimmunediseases, amyotrophic lateral sclerosis (ALS), oculopharyngeal musculardystrophy (OPMD), myotonic dystrophy (MD), Duchenne muscular dystrophy(DMD), polymyositis (PM), dermatomyositis (DM), inclusion body myositis(IBM), and other muscle disorders.

Inflammatory mechanisms have been proposed as important mediators in thepathogenetic cascade of Alzheimer's disease (McGeer P L, McGeer E G. Theinflammatory response system of brain implications for the therapy ofAlzheimer and other neurodegenerative diseases. Brain Res. Rev., 1995;21: 195-218). In the study by in't Veld et al. (the New England Journalof Medicine, 2001; 345, 1515), they followed almost 7000 person at riskof Alzheimer's disease for nearly seven years. Their results suggestedthat NSAIAs can reduce the relative risk for those whose cumulative useof NSAIAs was at least two years and two or more years before the onsetof dementia. If the neuroprotective capacity of NSAIAs ceases in theyears just before the onset of dementia, then these compounds wouldoffer no protection against progression among most persons with theprodromal stage of diseases. We believe that the reason for this is thatthe tissues around the damaged nerve cells will form scars to protectthe nerve cells from damaging farther. Most of NSAIAs have very lowbrain-blood and nerve cell barriers penetration rate and cannotpenetrate the scar barrier. These pro-drugs in this invention have veryhigh skin, blood-brain, nerve cell membrane, and scar barrierspenetration rates and are very promising agents for the treatment ofAlzheimer's disease, Parkinson's diseases, and other progressiveneurodegenerative diseases.

These pro-drugs may help the patients with a spinal cord injury in whichthe healing is stopped by the protected scars around the injured spinalcord. Most NSAIAs cannot penetrate the scar barrier in a therapeuticeffective amount, but the pro-drugs in this invention can penetrate thescar barrier, have anti-inflammatory activity, and can help woundhealing.

NSAIAs are not very effective for treatment of the conditions describedabove or have serious side effects because they cannot penetrate thecell membrane, especially the brain cells and nerve cells, veryeffectively and stay the general circulation too long, thus most ofdrugs will be metabolized by intestinal mucosa, liver, kidney, and lungbefore they reach the “site of action.” This situation not only producesvery low pharmacologic effect, but also causes toxic burden onintestinal mucosa, liver, kidneys, lungs, and other parts of the body.These pro-drugs in this invention penetrate skin, brain-blood, braincells, nerve cells and other membranes barriers very well and they havehundreds of times more potency than the parent drugs, only a few tenthsor hundredths of the normal drug dosage is needed and much less sideeffects will be caused. This will benefit not only transdermal drugdelivery, but also any other drug delivery system (such as oral,subcutaneous, intravenous, inhalation, and nasal) and can treat manyconditions better than they can be treated by their respective parentdrugs and even some conditions which cannot be treated by theirrespective parent drugs.

The compounds of the general formula (1, 2a, 2b, 2c, or 2d) “Structure1, 2a, 2b, 2c, or 2d” indicated above can be prepared from NSAIAs, byreaction with N,N′-Dicyclohexylcarbodiimide,N,N′-Diisopropylcarbodiimide or other coupling reagents to formanhydrides, then react with suitable alcohols, thiols, or amines.

The compounds of the general formula (1, 2a, 2b, 2c, or 2d) “Structure1, 2a, 2b, 2c, or 2d” indicated above can be prepared from metal salts,organic base salts, or immobilized base salts of NSAIAs with suitablehalide compounds.

Transdermal therapeutic application systems of a compound of the generalformula (1, 2a, 2b, 2c, or 2d) “Structure 1, 2a, 2b, 2c, or 2d” or acomposition comprising of at least one compound of the general formula(1, 2a, 2b, 2c, or 2d) “Structure 1, 2a, 2b, 2c, or 2d” as an activeingredient, can be used for treating any NSAIAs-treatable conditions andany conditions described in this invention in humans or animals. Thesesystems can be a bandage or a patch comprising of one activesubstance-containing matrix layer and an impermeable backing layer. Themost preferable system is an active substance reservoir, which has apermeable bottom facing the skin. By controlling the rate of release,this system enables NSAIAs to reach constantly optimal therapeutic bloodlevels to increase effectiveness and reduce the side effects of NSAIAs.These systems can be worn on the wrist, ankle, arm, leg, or any part ofbody.

Advantageous Effects

NSAIAs are not very effective for treatment of the conditions describedin this invention or have serious side effects because they cannotpenetrate the cell membrane, especially the brain cells and nerve cells,very effectively and stay in the general circulation too long, thus mostof drugs will be metabolized by intestinal mucosa, liver, kidneys, andlungs before they reach the “site of action.” This situation not onlyproduces very low pharmacologic effect, but also causes toxic burden onintestinal mucosa, liver, kidneys, lungs and any other parts of thebody. These pro-drugs in this invention penetrate skin, brain-blood,brain cells, nerve cells, scars and other membranes barriers very well,and they have hundreds of times more potency than the parent drugs, onlya few tenths or hundredths of the normal drug dosage is needed and muchless side effects will be caused. This will benefit not only transdermaldrug delivery, but also any other drug delivery system (such as oral,subcutaneous, intravenous, inhalation, and nasal) and can treat manyconditions that cannot be treated by their parent drugs. These pro-drugscan be administered not only transdermally, but also orally (they willnot hurt stomach because they cannot penetrate the wall of the stomach)for any type of medical treatment and should avoid most of the sideeffects of NSAIAs, most notably GI disturbances such as dyspepsia,gastroduodenal bleeding, gastric ulcerations, and gastritis. Anothergreat benefit of transdermal administration of these pro-drugs is thatadministering medication, especially to children, will be much easier.

DESCRIPTION OF DRAWINGS

FIG. 1 depicts the chemical structure for Structure 1, FIGS. 2A, 2B, 2C,and 2D depict the chemical structures of Structure 2a, Structure 2b,Structure 2c, and Structure 2d, respectively.

BEST MODE Preparation of DiethylaminoethylAcetylsalicylate.Acetylsalicylic Acid Salt

108 g of 2-acetylsalicyclic acid was dissolved in 1000 ml of chloroform.The mixture was cooled to 5° C. 103 g of 1,3-Dicyclohexylcarbodiimidewas added into the mixture. The mixture is stirred for 2 h at RT. Thesolid waste is removed by filtration and washed with chloroform (3×300ml). 59 g of diethylaminoethanol were added into the reaction mixture.The mixture was stirred for 3 hours at RT. The organic solution wasevaporated off. After drying, it yielded 220 g of the desired product(96%). Elementary analysis: C₂₄H₂₉NO₈; MW: 459.18. Calculated % C,62.73; H, 6.36; N, 3.05; O, 27.86. Found. % C, 62.70; H, 6.40; Cl: N,3.01; O, 27.90.

Mode for Invention Preparation of 1-piperidinepropyl2[(2,6-dichlorophenyl)amino]benzene acetate.AcOH

31.8 g (0.1 mol) of sodium 2[(2,6-dichlorophenyl)amino]benzene acetatewas suspended in 180 ml of chloroform. 28.6 g (0.1 mol) of1-piperidinepropyl bromide.HBr was added into the mixture and themixture was stirred for 5 hours at RT. The mixture is washed with 5%Na₂CO₃ (1×300 ml) and water (3×100 ml). The mixture is dried overanhydrous Na₂SO₄. Sodium sulfate was removed by filtration and washedwith chloroform (3×50 ml). 6 g of acetic acid was added into thesolution. The solution is concentrated in vacuo to 100 ml. Then 300 mlof hexane was added into the solution. The solid product was collectedby filtration and washed with hexane (3×100 ml). After drying, ityielded 40 g of the desired product (86%). Elementary analysis:C₂₄H₃₀Cl₂N₂O₄; MW: 481.43 Calculated % C, 59.88; H, 6.28; Cl, 14.73; N,5.82; O, 13.29. Found. % C, 59.83; H, 6.32; Cl, 14.71; N, 5.79; O,13.35.

Preparation of 3-piperidinemethyl 2-(ρ-isobutylphenyl)propionate.AcOH

60 g of Polymer-bound triethylamine (3 mmol/g, 100-200 mesh) wassuspended in 500 ml of chloroform. 20.6 g (0.1 mol) of2-(ρ-isobutylphenyl)propionic acid was added into the mixture withstirring. 39 g (0.15 mol) of 3-piperidinemethyl bromide.HBr was addedinto the mixture and the mixture was stirred for 10 hours at RT. Thepolymer is removed by filtration and washed with acetone (3×50 ml). 300ml of 5% Na₂CO₃ was added into the solution with stirring. The mixtureis stirred for 30 min. The chloroform solution is washed with water(3×100 ml) and dried over Na₂SO₄. Sodium sulfate is removed byfiltration and washed with chloroform (3×100 ml). 6 g of acetic acid wasadded into the mixture. The solution is concentrated in vacuo to 100 ml.Then 300 ml of hexane was added into the solution. The solid product wascollected by filtration and washed with hexane (3×100 ml). After drying,it yielded 35 g of the desired product (96%). Elementary analysis:C₂₁H₃₃NO₄; MW: 363.49 Calculated % C, 69.39; H, 9.15; N, 3.85; O, 17.61.Found. % C, 69.35; H, 9.18; N, 3.83; O, 17.64.

INDUSTRIAL APPLICABILITY

The pro-drugs of the general formulas (1, 2a, 2b, 2c, or 2d) “Structure1, 2a, 2b, 2c, or 2d” are superior to NSAIAs. They can be usedmedicinally in treating any NSAIAs-treatable conditions in humans oranimals. They can be used also for treating and preventing diabetes(type I & II), abnormal blood glucose and lipid levels, stroke, heartattack, and other heart and vascular diseases Alzheimer's diseases,Parkinson's diseases and other neurodegenerative diseases, psoriasis,discoid lupus erythematosus, systemic lupus erythematosus (SLE),autoimmune hepatitis, scleroderma, Sjogren's syndrome, rheumatoidarthritis, polymyositis, scleroderma, Hashimoto's thyroiditis, juvenilediabetes mellitus, Addison disease, vitiligo, pernicious anemia,glomerulonephritis, and pulmonary fibrosis, multiple sclerosis (MS),Crohn's disease, and other autoimmune diseases, amyotrophic lateralsclerosis (ALS), oculopharyngeal muscular dystrophy (OPMD), myotonicdystrophy (MD), Duchenne muscular dystrophy (DMD), polymyositis (PM),dermatomyositis (DM), inclusion body myositis (IBM), and other muscledisorders, hemorrhoids, inflamed hemorrhoids, post irradiation(factitial) proctitis, chronic ulcerative colitis, cryptitis, otherinflammatory conditions of the anorectum, and pruritus ani, prostatitis,prostatocystitis, autoimmune liver inflammation, autoimmune kidneyinflammation, vein inflammation and other inflammations, spinal cordinjuries, scars, breast cancer, colon-rectum cancer, oral cancer, lungand other respiratory system cancers, skin cancer, uterus cancer,genital cancer, urinary organs cancers, leukemia and other blood andlymph tissues cancers and other cancers, and many other conditions.These pro-drugs can be administered transdermally without the help ofskin penetration enhancers.

What is claimed is:
 1. A compound of the general formula (2b) “Structure 2b”,

wherein: R represents O; R₁ represents CH₂; R₂ represents CH₂; R₄ represents H, C₁₋₁₂ alkyl, C₁₋₁₂ alkyloxy, C₂₋₁₂ alkenyl, C₁₋₁₂ alkyl halide, or C₂₋₁₂ alkynyl aryl, or heteroaryl; X represents a bond; HA represents none or a pharmaceutically acceptable acid; Ary- represents

wherein, R_(x) represents H, CH₃, CH₃O, HO, CH₃CH₂, CF₃, CHF₂, CH₂F, Cl, F, Br, or F; Y, Y₁, Y₂, Y₃, or Y₄ represents independently H, HO, CH₃COO, R_(y)COO, HS, NO₂, CN, CH₃COS, NH₂, CH₃CONH, R_(y)CONH, CH₃, CH₃CH₂, C₃H₇, C₄H₉, CH₃O, CH₃CH₂O, C₃H₇O, Cl, F, Br, I, CH₃S, CHF₂O, CF₃O, CF₃CF₂O, C₃F₇O, CF₃, CF₃CF₂, C₃F₇, C₄F₉, CH₃SO₂, R_(y)SO₂, CH₃SO, R_(y)SO, CH3CO, or CH₃CH₂CO; R_(y) represents H, C₁₋₁₂ alkyl, C₁₋₁₂ alkyloxy, C₂₋₁₂ alkenyl, C₁₋₁₂ alkyl halide, C₂₋₁₂ alkynyl, aryl, or heteroaryl; Ary- may be achiral or chiral; and if Ary- is chiral, it may have one or more chiral centers and may be a single (R) or (S) enantiomer or a mixture of (R) and (S) enantiomers.
 2. A pharmaceutical composition comprising a compound according to claim 1 and a pharmaceutically acceptable excipient.
 3. The pharmaceutical composition according to claim 2, wherein the pharmaceutical composition is formulated in the form of a solution, spray, emulsion, pill, or tablet.
 4. The pharmaceutical composition of claim 2, which is for transdermal therapeutic application.
 5. The pharmaceutical composition of claim 2, consisting of a compound according to claim 1 and water.
 6. A transdermal therapeutic application system comprising a compound of claim 1 and an active substance reservoir.
 7. The compound according to claim 1, wherein HA is HCl, HBr, HF, HI, HOAc, or citric acid.
 8. The transdermal therapeutic application system according to claim 6, further comprising a bandage or a patch comprising one active substance-containing matrix layer and an impermeable backing layer.
 9. The transdermal therapeutic application system according to claim 6, wherein the active substance reservoir comprises a permeable bottom facing the skin. 